LOCUS       sequence-135588-        8651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the NCAM1, isoform 1.0.1 
            protein, C-terminally fused to alkaline phosphatase + 6X histidine 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8651)
  AUTHORS   Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray 
            TA, Kay JN, Wojtowicz WM
  TITLE     An extracellular biochemical screen reveals that FLRTs and Unc5s 
            mediate neuronal subtype recognition in the retina.
  JOURNAL   Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149.
  PUBMED    26633812
REFERENCE   2  (bases 1 to 8651)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Ncam1_1.0.1-AP-His
FEATURES             Location/Qualifiers
     source          1..8651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..633
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          748..1573
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     CDS             1650..1661
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5231..5248
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5274..5395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5311..5330)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5365..5384
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5433..5450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5604..6192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5684..5703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6363..7154,7155..7223))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6986..7005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7224..7328)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7396..7414
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7452..7474)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7476..7496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7481..7811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7662..7797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7724..7743
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7948..7967
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8034..8489)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8171..8192)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8383..8402
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8615..8637
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8629..8646
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8630..8646
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc
      781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc
      841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
      901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
      961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta
     1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc
     1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt
     1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc
     1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac
     1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt
     1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta
     1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg
     1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg
     1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg
     1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tgctgcgaac
     1621 taaggatctc atctggactt tgtttttcct gggaactgca gtttccctgc aggtagatat
     1681 tgttcccagc caaggagaaa tcagcgttgg agagtccaaa ttcttcctgt gtcaagtggc
     1741 aggagatgcc aaagataagg acatctcctg gttctccccc aatggggaga agctgagccc
     1801 aaaccagcag cggatctcag tggtgtggaa tgatgacgac tcctctaccc tcaccatcta
     1861 caacgccaac atcgacgatg ccggcatata caagtgtgtg gtcacggctg aggacggcac
     1921 ccagtctgag gccactgtca acgtgaagat cttccagaag ctcatgttca agaatgcacc
     1981 aaccccacag gagtttaagg aaggggagga tgctgtgatt gtctgtgatg tggtcagctc
     2041 cctgcctcca accatcatct ggaaacacaa aggccgagat gtcattctga aaaaagacgt
     2101 ccggttcata gtcctgtcca acaactacct gcagatcagg ggcatcaaga aaacagatga
     2161 gggtacttac cgctgtgagg gcaggatcct ggcccgcggg gaaatcaact tcaaggacat
     2221 tcaggtcatt gtgaatgtac cacccactgt ccaggccaga cagagcatcg tgaatgccac
     2281 tgccaacctg ggccagtctg tcaccctggt gtgtgatgcc gatggcttcc cagagcccac
     2341 catgagctgg acaaaggatg gggaacccat agagaacgag gaggaagatg acgagaagca
     2401 catcttcagt gatgacagct ccgagctgac catcaggaat gtggataaaa acgacgaggc
     2461 cgaatacgtc tgcatcgcag agaacaaggc tggcgagcag gatgcctcca tccacctcaa
     2521 ggtctttgca aagcccaaaa tcacctatgt agagaatcag acggccatgg aactagagga
     2581 gcaagtcact ctgacctgtg aagcctccgg agaccccatc ccctccatca cctggagaac
     2641 atccacccgg aacatcagca gtgaagaaaa ggcatcgtgg actcgaccag agaagcaaga
     2701 gactctggat gggcacatgg tggttcgcag ccatgctcgt gtgtcctcct tgaccctgaa
     2761 gagcatccag tacacagatg ctggagaata catctgcact gccagcaaca ccatcggcca
     2821 ggattcccag tccatgtacc tcgaagttca atatgctccc aagctccagg gccctgtggc
     2881 tgtgtacact tgggaaggga accaggtgaa catcacctgt gaggtcttcg cctacccaag
     2941 tgccacaatc tcctggttcc gagatggtca gttgctgcca agctccaact acagcaatat
     3001 caagatctac aacaccccat ctgctagcta tctggaggtg acccctgatt cagaaaatga
     3061 ctttggaaac tacaactgta cagcagtgaa ccgtattgga caggagtcct tggaattcat
     3121 ccttgttcaa gcagacacac cgtcttctcc atccattgac cgggtggaac catactccag
     3181 cacagcgcag gtgcagtttg atgagccaga ggccacaggg ggcgtgccca ttctcaagta
     3241 caaggctgag tggaagtcgc tgggcgaaga atcctggcat ttcaagtggt atgatgccaa
     3301 agaagccaac atggaaggca ttgtcaccat catgggcctg aaacctgaga cgaggtactc
     3361 ggtacgactg gcggccctca acggcaaggg gctgggcgag atcagtgcag ccactgagtt
     3421 caagacacag ccagtccggg aacccagtgc acccaagctg gaagggcaga tgggagagga
     3481 cgggaactcc atcaaggtga acctgatcaa gcaggatgac ggaggctccc ccatcagaca
     3541 ctatctggtc aagtacagag cgaagctcgc ctctgagtgg aaaccggaaa tcaggctccc
     3601 atccggcagt gaccacgtca tgctcaagtc cctggactgg aacgcagagt atgaagtcta
     3661 tgtggtagct gaaaaccagc aaggaaaatc caaggcagct cactttgtgt tcaggacctc
     3721 agcccagccc acggccatcc cagccactag tggatcttcc ggaatcatcc cagttgagga
     3781 ggagaacccg gacttctgga accgcgaggc agccgaggcc ctgggtgccg ccaagaagct
     3841 gcagcctgca cagacagccg ccaagaacct catcatcttc ctgggcgatg ggatgggggt
     3901 gtctacggtg acagctgcca ggatcctaaa agggcagaag aaggacaaac tggggcctga
     3961 gatacccctg gccatggacc gcttcccata tgtggctctg tccaagacat acaatgtaga
     4021 caaacatgtg ccagacagtg gagccacagc cacggcctac ctgtgcgggg tcaagggcaa
     4081 cttccagacc attggcttga gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg
     4141 caacgaggtc atctccgtga tgaatcgggc caagaaagca gggaagtcag tgggagtggt
     4201 aaccaccaca cgagtgcagc acgcctcgcc agccggcacc tacgcccaca cggtgaaccg
     4261 caactggtac tcggacgccg acgtgcctgc ctcggcccgc caggaggggt gccaggacat
     4321 cgctacgcag ctcatctcca acatggacat tgacgtgatc ctaggtgggg gccgaaagta
     4381 catgtttcgc atgggaaccc cagaccctga gtacccagat gactacagcc aaggtgggac
     4441 caggctggac gggaagaatc tggtgcagga atggctggcg aagcgccagg gtgcccggta
     4501 tgtgtggaac cgcactgagc tcatgcaggc ttccctggac ccgtctgtga cccatctcat
     4561 gggtctcttt gagcctggag acatgaaata cgagatccac cgagactcca cactggaccc
     4621 ctccctgatg gagatgacag aggctgccct gcgcctgctg agcaggaacc cccgcggctt
     4681 cttcctcttc gtggagggtg gtcgcatcga ccatggtcat catgaaagca gggcttaccg
     4741 ggcactgact gagacgatca tgttcgacga cgccattgag agggcgggcc agctcaccag
     4801 cgaggaggac acgctgagcc tcgtcactgc cgaccactcc cacgtcttct ccttcggagg
     4861 ctaccccctg cgagggagct ccatcttcgg gctggcccct ggcaaggccc gggacaggaa
     4921 ggcctacacg gtcctcctat acggaaacgg tccaggctat gtgctcaagg acggcgcccg
     4981 gccggatgtt accgagagcg agagcgggag ccccgagtat cggcagcagt cagcagtgcc
     5041 cctggacgaa gagacccacg caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc
     5101 gcacctggtt cacggcgtgc aggagcagac cttcatagcg cacgtcatgg ccttcgccgc
     5161 ctgcctggag ccctacaccg cctgcgacct ggcgcccccc gccggcacca ccgacgccgc
     5221 gcacccgggt catcatcacc atcaccatta atctagagat cttattaaag cagaacttgt
     5281 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5341 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5401 tctggtcgac tctagctaga ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5461 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5521 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5581 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5641 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5701 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5761 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5821 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5881 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5941 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6001 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6061 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6121 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6181 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6241 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6301 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6361 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6421 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6481 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6541 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6601 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6661 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6721 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6781 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6841 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6901 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6961 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7021 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7081 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7141 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7201 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7261 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7321 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     7381 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     7441 gacggtgaaa acctctgaca catgcagctc ccggaggaat gtgtgtcagt tagggtgtgg
     7501 aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7561 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     7621 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     7681 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     7741 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     7801 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cgtcccggag acggtcacag
     7861 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7921 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     7981 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     8041 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     8101 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     8161 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     8221 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     8281 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     8341 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     8401 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     8461 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     8521 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     8581 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     8641 gccagtgaat t
//
